This invention relates generally to cell biology, virology and medicine. In particular, the invention is directed to chimeric polypeptides, e.g., bispecific antibodies, comprising a chemokine receptor binding domain and a T cell surface polypeptide, a cell toxin, or a cell toxin binding domain, nucleic acids that encode them, and methods of making and using them. The chimeric polypeptides of the invention can include, be bound to, or attached to, a cell toxin. The invention is also directed to making and using pharmaceutical compositions, for example, for the treatment of immunological (e.g., autoimmune) disorders and for the targeted elimination of cells, e.g., T lymphocytes and other cells latently infected with a primate immunodeficiency virus, such as a human immunodeficiency virus, e.g., HIV-1. The pharmaceutical compositions and methods of the invention can be used for the treatment, prevention and/or alleviation of inflammatory joint and renal diseases, inflammatory bowel diseases, multiple sclerosis, skin diseases, diabetes or transplant rejection.
Immunological diseases/disorders, like autoimmune diseases, inflammation disorders as well as infectious diseases are not only increasing but represent substantial threats to global health. For example, in Germany, about 1% of the population suffer from the autoimmune disease rheumatoid arthritis. In addition, there is a number of other joint diseases also leading to arthritis. Currently, three groups of drugs-non-steroidal anti-rheumatics, cortisone preparations and second-line agents- and TNFxcex1 blocking agents are used for treating inflammatory joint diseases. Up to now, the therapy has focused on the local injection of cortisone preparations in combination with a systemic administration of anti-phlogistics or second-line agents.
Non-steroidal anti-rheumatics have a mild analgetic and anti-inflammatory effect, but they have many side effects when applied frequently (e.g. gastric ulcers, nephroses). In high dosages, cortisone preparations have a strong decongestant and analgetic effect, however, leading to a quick relapse after discontinuation of the therapy. Moreover, cortisone preparations cannot stop the destruction process of the joint disease. A long-term therapy with cortisone usually entails severe side effects, such as infections, Cushing""s phenomenon, osteoporosis, parchment-like skin, metabolic and hormonal disorders. The local injection of cortisone also has the essential disadvantage that the activity of the migrated white blood cells is only reduced. As the infiltrating cells are not destroyed, a quick relapse occurs after discontinuation of the therapy. As mentioned above, the same applies to the systemic application. Rarely, inflammation due to the irritative effect of cortisone crystals is aggravated after injection of cortisone. The duration of effect of a cortisone injection varies tremendously and ranges from primary ineffectiveness to a duration of effect of several weeks.
In rheumatology, second-line agents are used to achieve a long-term suppression of the inflammation and a reduction in cortisone preparations. Due to the considerable toxicity (e.g., allergies, infections, malignant diseases, renal insufficiency, blood pressure crises, pulmonary diseases) it is necessary for medical specialists to attend closely to the patients. After beginning treatment, no therapeutic effect may be apparent for the first three months. Currently, there are 4 or 5 of such second-line agents at disposal, which are used individually at first or are combined if the therapy is not effective. Mostly, there is hardly anything known about the mode of action of second-line agents. It is not yet entirely clear whether the application of second-line agents can diminish the destruction of the joint.
In recent years, a new group of substances has been introduced into the treatment of rheumatoid arthritis, which is based on the blocking of cell signal substances, particularly TNFxcex1, by means of monoclonal antibodies or soluble receptor constructs.
In addition, there are patients that do not respond to currently available therapies. In other cases, the conventional therapy has to be stopped due to intolerable side effects.
A similar situation exists for many other inflammatory and autoimmune diseases like inflammatory renal diseases, inflammatory bowel diseases, multiple sclerosis and transplant rejection, where current treatments have many limitations. For example, agents used in inflammatory and autoimmune diseases include anti-inflammatory and immunosuppressive agents like azathioprine, cyclophosphamide, glucocorticoids like prednisone; immunosuppressants like cyclosporin A, Tacrolimus (FK506), Sirolimus (Rapamycin); and protein drugs like calcineurin, beta-interferon, anti-TNF alpha monoclonal antibodies (remicade). These agents show general immunomodulating effects and therefore efficacy and side effects profiles can pose severe limitations for the treatment options; see, e.g., Harrison""s Principles of Internal Medicine, eds. Fauci et al., 14th edition, McGraw-Hill publisher.
Inflammatory bowel diseases, such as Crohn""s disease, ulcerative colitis, are treated with the anti-inflammatory agents sulfazsalazine (Azulfidine) and glucocorticoids, like prednisone and, in selected cases, with TNF-xcex1 blocking agents. In ulcerative colitis immunosuppressive therapy with drugs such as azathioprine is well established, in severely ill patients the potent immunosuppressive agent cyclosporine is used (see, e.g., Harrison""s Principles of Internal Medicine, eds. Fauci et al., 14th edition, McGraw-Hill publisher). In many cases no sufficient reduction of disease activity is achieved with current drugs, such that even surgical intervention is sometimes necessary.
Inflammatory renal diseases (nephritis) are treated with e.g. glucocorticoids, alkylating agents and/or plasmapheresis. Additional diseases with similar treatment options include systemic lupus erythematosus (SLE), Sjogren""s syndrome, polymyositis, dermatomyositis, mixed connective tissue disease, anti-phospholipid-antibody syndrome.
For some of these diseases, few therapeutic options have been available up to now. All these diseases share an inflammatory component. However, the inflammatory component cannot be sufficiently suppressed by the currently available drugs. For some drugs, e.g. alkylating agents a maximal lifetime dose per patient cannot be exceeded.
Transplant rejection is treated using immunosuppressive agents including azathioprine, mycophenolate mofetil, glucocorticoids, cyclosporine, Tacrolimus (FK506), Sirolimus (Rapamycin). A combination of steroids and a low dose of mouse monoclonal antibody OKT3 binding to CD3 on T-cells is used to anergize and deplete T-cells. Therapy is continued using immunosuppressants like cyclosporine. Mouse anti-human antibodies (MAHAs) have common side effects and limit the use of OKT3 (Fauci et al., supra, pp. 2374-2381).
Approaches to treat multiple sclerosis include treatments which effect the overall immune system like anti-inflammatory agents including azathioprine, cyclophosphamide, prednisone, corticosteroids, cyclosporin A, calcineurin, Rapamycin, beta-interferon (see, e.g., Fauci et al., supra, pp. 2415-2419; Wang (2000) J. Immunol. 165:548-557). In addition, a number of non-specific treatments are administered that may improve the quality of life including physical therapy and psycho-pharmacological agents. None of the treatment options mentioned above has a curative effect. Even the most promising compound, xcex2-interferon, leads only to a slower disease progression, while exhibiting significant side effects.
Furthermore, human immunodeficiency virus-type 1 (HIV-1), the most common cause of AIDS, has infected more than 50 million individuals (including those who have died), and the rate of new infections is estimated at nearly 6 million per year (AIDS Epidemic Update: December 1999 (UNAIDS, Geneva, 1999), www.unaids.org). Equally disturbing are the uncertainties of the epidemic to come. Although sub-Saharan Africa remains the global epicenter, rates of infection have increased in recent times in the former Soviet Union and parts of south and southeast Asia, including India and China, where literally hundreds of millions of individuals are potentially at risk. In the United States, new waves of infection have been recognized in women, minorities, and younger generations of gay men. Combination antiretroviral therapy has afforded many people clinical relief, but the costs and toxicities of treatment are substantial, and HIV-1 infection remains a fatal disease. Moreover, the vast majority of infected people worldwide do not have access to these agents. Thus, although the demographics (and, in some instances, the natural history) of AIDS have changed, the epidemic is far from over; instead, it is evolving, expanding, and posing ever greater challenges.
Human immunodeficiency virus (HIV) cannot enter human cells unless it first binds to two key molecules on the cell surface, CD4 and a co-receptor. The co-receptor that is initially recognized is chemokine receptor 5 (CCR5). Later in the life cycle of the virus, another chemokine receptor, CXCR4, becomes the co-receptor for HIV-1; see, e.g., D""Souza, Nature Med. 2:1293 (1996); Premack, Nature Med. 2:1174; Fauci, Nature 384:529 (1996).
The HIV-1 strains that cause most transmissions of viruses by sexual contact are called M-tropic viruses. These HIV-1 strains, also known as NSI primary viruses, can replicate in primary CD4+ T-cells and macrophages and use the chemokine receptor 5 (CCR5), and, less often, CCR3, as their entry co-receptor. The T-tropic viruses, sometimes called SI primary, can also replicate in primary CD4+ T-cells, but can, in addition, infect established CD4+ T-cell lines in vitro, which they do via the chemokine receptor CXCR4 (fusin). Many of these T-tropic strains can use CCR5 in addition to CXCR4, and some can enter macrophages via CCR5, at least under certain in vitro conditions; see, e.g., D""Souza, Nature Med. 2, 1293 (1996); Premack, Nature Med. 2, 1174; Fauci, Nature 384, 529 (1996).
Whether other co-receptors contribute to HIV-1 pathogenesis is unresolved, but the existence of another co-receptor for some T-tropic strains can be inferred from in vitro studies. Because M-tropic HIV-1 strains are implicated in about 90% of sexual transmissions of HIV, CCR5 is the predominant co-receptor for the virus in patients; transmission (or systemic establishment) of CXCR4-using (T-tropic) strains is rare (D""Souza, Nature Med. 2, 1293 (1996); Premack, Nature Med. 2, 1174; Fauci, Nature 384, 529 (1996), Paxton, Nature Med. 2, 412 (1996); Liu, Cell 86, 367 (1996); Samson, Nature 382, 722 (1996); Dean, Science 273, 1856 (1996); Huang, Nature Med. 2, 1240 (1996)). However, once SI viruses evolve in vivo (or if they are transmitted), they are especially virulent and cause faster disease progression; see, e.g., D""Souza, Nature Med. 2, 1293 (1996); Premack, Nature Med. 2, 1174; Fauci, Nature 384, 529 (1996), Schuitemaker, J. Virol. 66, 1354 (1992); Connor, J. Virol. 67, 1772 (1993); Richman, J. Infect. Dis. 169, 968 (1994); R. I. Connor (1997) J. Exp. Med. 185:621; Trkola, Nature 384, 184 (1996).
The numbers and identity of co-receptor molecules on target cells, and the ability of HIV-1 strains to likely enter cells via the different co-receptors, seem to be critical determinants of disease progression. These factors are major influences on both host- and virus-dependent aspects of HIV-1 infection. For example, a homozygous defect (delta 32) in CCR5 correlates strongly with resistance to HIV-1 infection in vivo and in vitro. Individuals who are heterozygous for a defective CCR5 allele are not protected against infection and have only a modestly slowed disease progression (Paxton, Nature Med. 2, 412 (1996); Liu, Cell 86, 367 (1996); Samson, Nature 382, 722 (1996); Dean, Science 273, 1856 (1996); Huang (1996) Nature Med. 2:1240).
However, other factors can influence the level of CCR5 expression on activated CD4+ T-cells and thereby affect the efficiency of HIV-1 infection in vitro (Trkola, Nature 384, 184 (1996); Bleul, Proc. Natl. Acad. Sci. U.S.A. 94, 1925 (1997)). For reasons that are not yet clear, the amount of CCR5 expression on the cell surface (as measured by MIP-1 binding) varies by 20-fold on CD4+ T-cells from individuals with two wild-type CCR5 alleles (Trkola, Nature 384, 184 (1996)). Staining with a CCR5-specific monoclonal antibody indicates a similar large variability (Wu, J. Exp. Med. 186:1373-81 (1997)). Such variation may far outweigh any effect of one defective allele for CCR5. The causes of this variation should be the subject of intensive studies, as they point to controllable factors that could increase resistance to disease.
Most primary, clinical isolates of primate immunodeficiency viruses use the chemokine receptor CCR5 for entry (see, e.g., Feng, Science 272, 872 (1996); Choe, Cell 85, 1135 (1996); Deng, Nature 381, 661 (1996); Dragic et al., Nature 381, p. 667; Doranz, Cell 85, 1149 (1996); Alkhatib, Science 272, 1955 (1996)). For most HIV-1 isolates that are transmitted and that predominate during the early years of infection, CCR5 is an obligate co-receptor, and rare individuals that are genetically deficient in CCR5 expression are relatively resistant to HIV-1 infection (see, e.g., Connor, J. Exp. Med. 185, 621 (1997); Zhang, Nature 383, 768 (1996); Bjxc3x6rndal, J. Virol. 71, 7478 (1997); Dean, Science 273, 1856 (1996); Liu, Cell 86, 367 (1996); Paxton, Nature Med. 2, 412 (1996); Samson, Nature 382, 722 (1996)). HIV-1 isolates arising later in the course of infection often use other chemokine receptors, frequently CXCR4, in addition to CCR5. Studies of chimeric envelope glycoproteins demonstrated that the third variable (V3) loop of gp 120 is a major determinant of which chemokine receptor is used as a viral entry co-receptor (see, e.g., Cocchi, Nature Med. 2, 1244 (1996); Bieniasz, EMBO J. 16, 2599 (1997); Speck, J. Virol. 71, 7136 (1997)). V3-deleted versions of gp120 do not bind CCR5, even though CD4 binding occurs at wild-type levels. Antibodies to the V3 loop interfere with gp120-CCR5 binding (see, e.g., Trkola, Nature 384, 184 (1996); Wu, Nature 384, 179 (1996); Lapham, Science 274, 602 (1996); Bandres, J. Virol. 72, 2500 (1998); Hill, Science 71, 6296 (1997)). These results support an involvement of the V3 loop in chemokine receptor binding.
Latency of HIV is established very early in the course of an infection, when M-tropic strains predominate. M-tropic strains depend on the presence of CCR5 on the target cell for infection. The importance of CCR5 as an essential co-receptor for M-tropic HIV-1 is emphasized by the fact that individuals lacking CCR5 due to a homozygous 32 base pair deletion (delta32) are highly resistant to HIV-1 infection. In contrast to other markers like CD4 or CD45RO, CCR5 is only present on a subset of lymphocytes and other cells that are prone to HIV-1 infection (Rottmann (1997) Am. J. Pathol. 151:1341-1351; Naif (1998) J. Virol. 72:830-836; Lee (1999) Proc. Natl Acad. Sci. USA 96:5315-5220).
Several approaches have been postulated to eliminate latent infected cells. One strategy is to drive the latently infected cells to virus production and subsequent cell death. In this context, one approach is IL-2 (or TNF-alpha or IL-6) administration in the presence of HAART until the viral reservoir is exhausted (Chun (1998) J. Exp. Med. 188, 83-91; Chun (1999) Nat. Med. 5, 651-655; Stellbrink (1999) Abstracts of the 6th Conference on Retroviruses and Opportunistic Infections (Foundation for Retrovirology and Human Health, Alexandria, Va.), abstr. 356. p. 135; Imamichi (1999) Abstracts of the 6th Conference on Retroviruses and Opportunistic Infections (Foundation for Retrovirology and Human Health, Alexandria, Va.), abstr. 358, p. 135). These cells are believed to die after activation. Whether the entire pool of latent infected cells can be exhausted is questionable.
Another strategy tried was to specifically kill latently infected cells based on gp-120 expression on the cell surface. Immunotoxins recognizing gp-120 have been proposed but failed for two reasons. The one construct tested in humans was a protein consisting of soluble CD4 linked to Pseudomonas aeroginosa exotoxin A (PE). The clinical results were disappointing due to dose-limiting hepatotoxicity without showing signs of efficacy and the program was terminated (Ashorn (1990) Proc. Natl Acad. Sci 87, 8889-8893; Berger (1998) Proc. Natl Acad. Sci. 95, 11511-11513). The second reason for failure was that latent infected cells do not express viral surface glycoproteins, e.g. gp-120 and gp-41. Thus, approaches targeting gp-120 or gp-41 for the elimination of latently infected cells cannot work.
Other approaches to eliminate latent infected cells are based on eliminating the entire CD4+ T-cell compartment (Berger (1998) Proc. Natl Acad. Sci. 95, 11511-11513), or the CD25-positive compartment (Bell (1993) Proc. Natl Acad. Sci. 90, 1411-1415), or the CD45RO memory cell compartment (McCoig (1999) Proc. Natl. Acad. Sci 96, 11482-11485). However these markers do not adequately include all potentially infected cells. Such cells also include, besides CD4-positive cells, macrophages, and non-hematopoietic cells.
In Wu, et al., WO 98/18826, an antibody directed against the mammalian (e.g. human) chemokine receptor 5 (CCR5) is described and said antibody is proposed in a method of inhibiting the interaction of cell bearing CCR5 with a potential ligand, like HIV. It is proposed that said method inhibits an HIV infection. Furthermore, treatment options for inflammatory diseases, autoimmune diseases and graft rejection are proposed. Yet, all these treatment options are based on the assumption that specific antibodies, like the immunoglobulin molecules themselves, or functional portions thereof, interfere with receptor-ligand interactions. However, whether these antibodies are capable of depleting the relevant cells is questionable. Furthermore, WO 98/18826 merely envisages the prevention of an interaction of HIV and the CCR5 receptor and thereby preventing an HIV infection.
Leukocytes, in particular T-cells, are believed to be the key regulators of the immune response to infective agents and are critical components for the initiation and maintenance of inflammatory processes, like inflammatory bowel disease inflammatory renal diseases, inflammatory joint disease, autoimmune disorders, like multiple sclerosis and arthritis, skin diseases, like psoriatic lesions, diabetes and in transplant rejection.
Thus, there exists a need for novel means and methods which can lead to the suppression of activated leukocytes involved in immunological pathologies, like autoimmune diseases, inflammation process and/or viral infections of immune cells. The present invention fulfills this and other needs.
The invention provides a chimeric polypeptide, e.g., a bispecific antibody, comprising a first polypeptide domain comprising at least one moiety that specifically binds to a chemokine receptor; and, a second polypeptide domain comprising at least one moiety that specifically binds to a T cell surface polypeptide or a cell toxin, or, a cell toxin. In one aspect of the invention, the chemokine receptor is a chemokine receptor 5 (CCR5), such as a human chemokine receptor 5 (CCR5). In one aspect, the moiety that specifically binds to the chemokine receptor 5 (CCR5) can comprise a RANTES (xe2x80x9cregulated on activation normal T cell expressed and secretedxe2x80x9d) polypeptide, or a fragment thereof capable of binding to a CCR5 receptor. Alternatively, the moiety that specifically binds to the CCR5 chemokine receptor can comprise a MIP-1xcex1, or a fragment thereof capable of binding to a CCR5 receptor. In another aspect, the moiety that specifically binds to the CCR5 chemokine receptor can comprise a MIP-1xcex2, a MCP-2, or a MCP-3 or a fragment thereof, capable of binding to the CCR5 receptor.
In one aspect of the chimeric composition of the invention, the moiety that specifically binds to the chemokine receptor comprises an IP-10 (CXCL-10) (see, e.g., Agostini (2001) Am. J. Pathol. 158:1703-1711; Flier (1999) J. Invest. Dermatol. 113:574-578), or a Mig (CXCL9) (see, e.g., Farber (1997) J. Leukoc. Biol. 61:246-257), or an I-TAC (CXCL11) chemokine ligand (see, e.g., Gasperini (1999) J. Immunol. 162:4928-4937), or a fragment thereof (see Table IV), capable of binding to the CXCR3 chemokine receptor.
In alternative aspects, the chemokine receptor is CXCR4 (see, e.g., Vila-Coro (1999) FASEB J. 13:1699-1710), CXCR5 (see, e.g., Legler (1998) J. Exp. Med. 187:655-660), CXCR6 (see, e.g., Luttichau (2001) Eur. J. Immunol. 31:1217-1220), CCR1 (see, e.g., Hesselgesser (1998) J. Biol. Chem. 273:15687-15692), CCR2 (see, e.g., Monteclaro (1997) J. Biol. Chem. 272:23186-23190), CCR3 (see, e.g., Dairaghi (1997) J. Biol. Chem. 272:28206-28209), CCR4 (see, e.g., Imai (1997) J. Biol. Chem. 272:15036-15042), CCR5 (see, e.g., Ganju (2000) J. Biol. Chem. 275:17263-17268), CCR6 (see, e.g., Baba (1997) J. Biol. Chem. 272:14893-14898), CCR7 (see, e.g., Kim (1999) Cell Immunol. 193:226-235), CCR8 (see, e.g., Roos (1997) J. Biol. Chem. 272:17251-17254), CCR9 (see, e.g., Norment (2000) J. Immunol. 164:639-648), CCR10 (see, e.g., Bonini (1997) DNA Cell Biol. 16:1249-1256), XCR1 (GPR5) (see, e.g., Shan (2000) Biochem. Biophys. Res. Commun. 268:938-941), or CX3CR1 (see, e.g., Combadiere (1998) Biochem. Biophys. Res. Commun. 253:728-732); see Table IV, which includes the corresponding chemokine ligands. The T cell surface polypeptide can comprise a CD3 polypeptide.
In one aspect, the chimeric composition of the invention comprises a cell toxin, or a fragment or domain thereof that remains toxic to cells. The cell toxin can comprise a Pseudomonas exotoxin, or toxic fragment thereof. The Pseudomonas exotoxin can comprise a PE38 exotoxin, a PE40 exotoxin or a PE37 exotoxin. Alternatively, the cell toxin can comprise a diptheria toxin. The cell toxin can be non-covalently or covalently, directly or indirectly, attached or associated with the chimeric composition. In one aspect, the toxin is cross-linked to the chimeric polypeptide. Alternatively, the toxin can comprise a recombinant fusion protein, as all or a portion of the chimeric polypeptide can comprise a recombinant protein, e.g., it can be a fusion protein. In one aspect, the moiety that specifically binds to a chemokine receptor comprises an antigen binding domain derived from an antibody that specifically binds to the chemokine receptor. The moiety that specifically binds to a T cell surface polypeptide can comprise an antigen binding domain derived from an antibody that specifically binds to the T cell surface polypeptide. The moiety that specifically binds to a cell toxin can comprise an antigen binding domain derived from an antibody that specifically binds to the cell toxin.
The invention also provides a recombinant fusion protein comprising a first polypeptide domain comprising at least one moiety that specifically binds to a chemokine receptor; and, a second polypeptide domain comprising at least one moiety that specifically binds to a T cell surface polypeptide or a cell toxin, or, a cell toxin.
The invention also provides a bispecific antibody comprising a first antigen binding domain that specifically binds to a chemokine receptor; and, a second antigen binding domain that specifically binds to a T cell surface polypeptide, a cell toxin, or a third antigen binding domain that specifically binds to or is linked to a T cell surface polypeptide or a cell toxin. The bispecific antibody is not limited to two binding domains. The T cell surface polypeptide can comprise a CD3 antigen.
In one aspect of the bispecific antibody of the invention, the bispecific antibody is a single chain antibody construct. The single chain antibody construct can comprise a VL and a VH domain capable of specifically binding the chemokine receptor and a VH and a VL domain capable of specifically binding a T cell surface polypeptide. In one aspect, the antigen binding domain that specifically binds to a chemokine receptor can comprise a murine anti-human CCR5 antibody MC-1. In one aspect, the VL and VH domains are arranged in the order VL(MC-1)-VH(MC-1)-VH(CD3)-VL(CD3). The VL(MC-1) domain can comprise an amino acid sequence as set forth in SEQ ID NO:12. The VH(MC-1) domain can comprise an amino acid sequence as set forth in SEQ ID NO:16. The VH(CD3) domain can comprise an amino acid sequence as set forth in SEQ ID NO:26. The VL(CD3) domain can comprise an amino acid sequence as set forth in SEQ ID NO:28. The amino acid sequence of the bispecific antibody can be encoded by a nucleic acid as set forth in SEQ ID NO: 17, or comprising an amino acid sequence as set forth in SEQ ID NO: 18.
In one aspect of the bispecific antibody, the second antigen binding domain specifically binds to a cell toxin, or, the second antigen binding domain specifically binds to another domain (e.g., an antibody) that can specifically bind a toxin (or a cell surface protein). Alternatively, the antibody is covalently bound (directly or indirectly) to a cell toxin. The antibody can be bound to a second antibody that binds to a CD3 antigen or a cell toxin.
The invention provides a nucleic acid encoding a chimeric polypeptide (e.g., a bispecific antibody) comprising a first polypeptide domain comprising at least one moiety that specifically binds to a chemokine receptor; and, a second polypeptide domain comprising at least one moiety that specifically binds to a T cell surface polypeptide or a cell toxin, or, a cell toxin.
The invention provides a vector comprising a nucleic acid encoding a chimeric polypeptide (e.g., a bispecific antibody) comprising a first polypeptide domain comprising at least one moiety that specifically binds to a chemokine receptor; and, a second polypeptide domain comprising at least one moiety that specifically binds to a T cell surface polypeptide or a cell toxin, or, a cell toxin.
The invention provides a transformed cell comprising a nucleic acid encoding a chimeric polypeptide (e.g., a bispecific antibody) comprising a first polypeptide domain comprising at least one moiety that specifically binds to a chemokine receptor; and, a second polypeptide domain comprising at least one moiety that specifically binds to a T cell surface polypeptide or a cell toxin, or, a cell toxin.
The invention provides a pharmaceutical composition comprising a chimeric polypeptide of the invention, a nucleic acid of the invention, or a vector of the invention, or a transformed cell of the invention; and, a pharmaceutically acceptable excipient.
The invention provides a kit comprising a chimeric polypeptide (e.g., a bispecific antibody) of the invention, a nucleic acid of the invention, a vector of the invention, a transformed cell of the invention, or a pharmaceutical composition of the invention. The kit can further comprise pharmaceutically acceptable excipients. The kits can further comprise instructions on the specific uses of the pharmaceuticals of the invention, as set forth herein. The kit can further comprise ancillary or other drugs, e.g., where the kit is intended to be used to treat HIV-1 (e.g., AIDS), drugs employed in HAART also can be included in the kit.
The invention provides a use of a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention) to prepare a pharmaceutical composition for the elimination of cells that are latently infected with a primate (e.g., human) immunodeficiency virus, e.g., HIV-1, or a lentivirus.
The invention provides a use of a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention) to prepare a pharmaceutical composition for the treatment of an immunological disorder, such as an autoimmune disease, an allergic disease, a skin disease, an inflammatory disease, diabetes, graft versus host disease and transplant rejections. In alternative aspects, the autoimmune disease is, e.g., multiple sclerosis, type I diabetes and rheumatoid arthritis. In alternative aspects, the skin disease is a skin inflammation, an atopic dermatitis and psoriasis. In alternative aspects, the inflammatory disease is an inflammatory joint disease, such as arthritis (e.g., chronic arthritis), an inflammatory renal disease and an inflammatory bowel disease.
The invention provides a method for eliminating a cell infected with a primate immunodeficiency virus comprising administering a composition comprising a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention), in amounts sufficient to kill the cell. In one aspect, the primate immunodeficiency virus is a human immunodeficiency virus, such as HIV-1. The cell can be infected (e.g., latently infected) with a pathogen, e.g., a virus, such as a primate immunodeficiency virus.
The invention provides a method for the treatment of a primate immunodeficiency virus comprising the following steps: (a) providing a pharmaceutical composition comprising a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention), (b) administering the pharmaceutical composition in amounts sufficient to treat the primate immunodeficiency virus. The treatment can further comprise administration of other drugs, e.g., those employed in HAART, or other treatments.
The invention provides a method for the treatment of an inflammatory renal disease comprising the following steps: (a) providing a pharmaceutical composition comprising a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention), (b) administering the pharmaceutical composition in amounts sufficient to treat the inflammatory renal disease.
The invention provides a method for the treatment of an allergic reaction comprising the following steps: (a) providing a pharmaceutical composition comprising a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention), (b) administering the pharmaceutical composition in amounts sufficient to treat the allergic reaction.
The invention provides a method for the treatment of an inflammatory bowel disease comprising the following steps: (a) providing a pharmaceutical composition comprising a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention), (b) administering the pharmaceutical composition in amounts sufficient to treat the inflammatory bowel disease.
The invention provides a method for the treatment of multiple sclerosis comprising the following steps: (a) providing a pharmaceutical composition comprising a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention), (b) administering the pharmaceutical composition in amounts sufficient to treat the multiple sclerosis.
The invention provides a method for the treatment of a skin disease comprising the following steps: (a) providing a pharmaceutical composition comprising a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention), (b) administering the pharmaceutical composition in amounts sufficient to treat the skin disease.
The invention provides a method for the treatment of diabetes comprising the following steps: (a) providing a pharmaceutical composition comprising a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention), (b) administering the pharmaceutical composition in amounts sufficient to treat the diabetes.
The invention provides a method for the treatment of a transplant rejection comprising the following steps: (a) providing a pharmaceutical composition comprising a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention), (b) administering the pharmaceutical composition in amounts sufficient to treat the transplant rejection.
The invention provides a method for the treatment of inflammatory joint disease comprising the following steps: (a) providing a pharmaceutical composition comprising a chimeric polypeptide (e.g., a bispecific antibody) or a nucleic acid of the invention (e.g., a vector of the invention), (b) administering the pharmaceutical composition in amounts sufficient to treat the inflammatory joint disease. The inflammatory joint disease can comprise arthritis, such as rheumatoid arthritis.
The invention provides a method of making a chimeric composition that can bind to a chemokine receptor and a cell toxin comprising the following steps: (a) providing a first polypeptide comprising at least one moiety that specifically binds to a chemokine receptor and at least one moiety that specifically binds to a second polypeptide comprising an antigen binding domain, wherein the antigen comprises a cell toxin, and a compound comprising a cell toxin; (b) contacting the first and second polypeptide with the compound in vivo or in vitro under conditions wherein the first polypeptide specifically binds to the second polypeptide, and the second polypeptide specifically binds to the compound, thereby making the chimeric composition.
The invention provides a method of making a chimeric composition that can bind to a chemokine receptor and a T cell surface antigen comprising the following steps: (a) providing a first polypeptide comprising at least one moiety that specifically binds to a chemokine receptor and at least one moiety that specifically binds to a second polypeptide comprising an antigen binding domain, wherein the antigen comprises a T cell surface antigen binding domain; (b) contacting the first polypeptide with the second polypeptide in vivo or in vitro under conditions wherein the first polypeptide specifically binds to the second polypeptide, thereby making a chimeric composition. The T cell surface antigen can comprise a CD3 antigen.
The chimeric composition can further comprise a cell toxin covalently bound to the chimeric composition. The cell toxin can be a truncated sequence, e.g., a domain, that remains toxic to the cell, e.g., a Pseudomonas exotoxin A (PE38).
The present invention relates to the use of an antibody and/or a chemokine construct which binds to a chemokine receptor. These compositions are used to eliminate cells that are latently infected with a primate immunodeficiency virus; accordingly, the invention is also directed to pharmaceuticals comprising these compositions. The invention is also directed to use of these compositions for the preparation pharmaceutical compositions. The pharmaceutical compositions and methods using these pharmaceuticals for the treatment, prevention and/or alleviation of inflammatory renal diseases, inflammatory joint diseases, inflammatory bowel diseases, multiple sclerosis, skin diseases, diabetes or transplant rejection.
Furthermore, the invention relates to antibody constructs and/or chemokine constructs, in particular, to constructs wherein said antibody construct comprises a binding site for a chemokine receptor 5 (CCR5) and a binding site for CD3, wherein said chemokine construct comprises RANTES (xe2x80x9cregulated on activation normal T cell expressed and secretedxe2x80x9d) and a toxin. The invention also describes polynucleotides encoding said antibody or chemokine constructs, and vectors and hosts comprising said nucleic acid molecules. Additionally, the present invention relates to compositions comprising said antibody constructs, chemokine constructs, polynucleotides, vectors and/or hosts. The composition can be a pharmaceutical composition. Described is also the use of antibody constructs, the chemokine constructs, the polynucleotides, the hosts and/or the vectors for the preparation of a pharmaceutical composition for treating, preventing and/or alleviating an immunological disorder or for eliminating latently infected cells, wherein said cells are infected with a primate immunodeficiency virus, like HIV-1.
The present invention also relates to a method for treating, preventing and/or alleviating an immunological disorder or for the elimination cells that are latently infected with a primate immunodeficiency virus, such as HIV-1. Furthermore, the invention provides for a kit comprising the compounds of the invention. The kit can also include instructions on the use of pharmaceuticals in the kit.
Several documents are cited throughout the text of this specification. Each of the documents cited herein (including any manufacturers specifications, instructions, etc.) and all publications, patents, patent applications, GenBank sequences and ATCC deposits, cited herein are hereby expressly incorporated by reference for all purposes.